Process for preparing an anti-cancer agent, 1-((4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine, its crystalline form and its salts

ABSTRACT

The present invention relates a new process to synthesize 1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine (AL3818). A stable crystalline form of Al3818 has been prepared. Salts and their crystalline forms of AL3818 have been also prepared. Anti-cancer and optometric activities of AL3818 and its salts have been further tested. New process has been outlined in Scheme I.

This application is a continuation of U.S. application Ser. No.15/659,510, filed Jul. 25, 2017, which is a divisional of U.S.application Ser. No. 15/143,630, filed May 2, 2016, which claims thebenefit of U.S. Provisional Applications 62/156,734, filed on May 4,2015, and 62/205,272, filed on Aug. 14, 2015. The foregoing applicationsare fully incorporated herein by reference in their entireties for allpurposes.

FIELD OF THE INVENTION

The present invention relates a new process to synthesize1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine(AL3818). A stable crystalline form of Al3818 has been prepared. Saltsand their crystalline forms of AL3818 have been also prepared.Anti-cancer and optometric activities of AL3818 and its salts have beenfurther tested.

BACKGROUND OF THE INVENTION

1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)-cyclopropanamine(AL3818) has been structurally disclosed in WO2008112407 as anangiogenesis inhibitor with few preparation methods.

SUMMARY OF THE INVENTION Abbreviations and Definitions

The following abbreviations are used and have the meaning below for easeof reference.

EtOH: ethanol, MeOH: methanol, IPA: isopropanol, EtOAc: ethyl acetate,RT: room temperature, DIPEA: diisopropylethylamine, DCM:Dichloromethane, DMF: N,N-dimethylformamide, DMAP:4-N,N-dimethylaminopyridine, MsCl: methanesulfonyl chloride, THF:tetrahydrofuran, TFA: trifluoroacetic acid, TEA: triethylamine, Pd/C:Palladium on active Carbon,eq: equivalent, g: gram, mg: milligram, ml: milliliter, min: minutes,bis=di: two or doubleDSC: differential scanning calorimetric, TGA: thermogravimetricanalysis, XRPD: X-ray powder diffraction, Exo: exotherm, Endo:endotherm.ALL: Acute Lymphocytic or Lymphoblastic Leukemias, CLL: ChronicLymphocytic or Lymphoblastic Leukemias, AML: Acute Myelogenous orMyeloid Leukemias, CML: Chronic Myelogenous or Myeloid Leukemias

The term “C₁-C₆alkyl”, as used herein, unless otherwise indicated,includes 1 to 6 saturated monovalent hydrocarbon radicals havingstraight or branched moieties, including, but not limited to, methyl,ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, and thelike.

The term “C₁-C₆alkoxy”, as used herein, unless otherwise indicated,includes —OC₁-C₆alkyl groups wherein C₁-C₆alkyl is as defined above,such as methoxy and ethoxy.

Invention Scope

The present invention relates a new process to synthesize1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine(AL3818) by condensing intermediate (X1) with (Y1) in a solvent at thepresence of KI or NaI, or intermediate (X2) with (Y2) in a solvent toform intermediate (Z) which is deprotected to give the final compound(AL3818) in Scheme I. A stable crystalline form of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamineand its salts as well as crystalline forms of salts have also beenprepared.

Wherein, R is selected from H and C₁-C₆alkoxy.

DESCRIPTION OF DRAWINGS

FIG. 1. DSC graph of a crystalline form of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 2. TGA graph of a crystalline form of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 3. XRPD graph of a crystalline form of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 4. DSC graph of a crystalline form of bishydrochloride acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 5. TGA graph of a crystalline form of bishydrochloride acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 6. XRPD graph of a crystalline form of bishydrochloride acid saltof1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 7. DSC graph of a crystalline form of bishydrochloridehydrate acidsalt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 8. TGA graph of a crystalline form of bishydrochloridehydrate acidsalt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 9. XRPD graph of a crystalline form of bishydrochloridehydrate acidsalt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 10. DSC graph of a crystalline form of bismaleic acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 11. TGA graph of a crystalline form of bismaleic acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 12. XRPD graph of a crystalline form of bismaleic acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 13. DSC graph of a crystalline form of succinic acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 14. TGA graph of a crystalline form of succinic acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 15. XRPD graph of a crystalline form of succinic acid salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropanamine

FIG. 16. Effect of AL3818 and its salts on human endometrial cancerIshikawa xenografted athymic mice

FIG. 17. Effect of AL3818 salts combined with Carboplatin(CBX)/Paclitaxel (Taxol) on human endometrial cancer Ishikawaxenografted athymic mice

FIG. 18. Effects of oral administration of AL3818 on laser-induced CNV

FIG. 19. Effects of AL3818 (0.15 mg/kg body weight) and intravitrealanti-VEGF antibody on laser-induced CNV

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates a new process to synthesize1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine(AL3818) by condensing intermediate (X1) with (Y1) in a solvent at thepresence of KI or NaI, or intermediate (X2) with (Y2) in a solvent toform intermediate (Z) which is deprotected to give the final compound(AL3818) in Scheme I.

Wherein, R is selected from H and C₁-C₆alkoxy, preferably selected fromH and —OMe;

The present invention relates to prepare a stable crystalline form of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine;

The present invention relates to prepare the salts or stable crystallinesalt forms of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine;

The present invention relates to bishydrochloride acid,bishydrochloridehydrate acid, bismaleic acid and succinic acid salt, andtheir stable crystalline salt forms or stable crystalline free base formof1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)-cyclopropanamine.

The present invention relates to prepare a pharmaceutical compositionthat comprises a stable crystalline form of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamineand a pharmaceutically acceptable carrier;

The present invention relates to prepare a pharmaceutical compositionthat comprises the salts or stable crystalline salt forms of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)-methyl)cyclopropanamineand a pharmaceutically acceptable carrier;

The present invention relates to a stable crystalline form or the saltsor stable crystalline salt forms of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)-cyclopropanaminefor a method in treating a neoplastic disease;

The present invention relates to a stable crystalline form or the saltsor stable crystalline salt forms of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine for use in the manufacture of a medicament for a methodin the treating a neoplastic disease;

The present invention relates to a stable crystalline form or the saltsor stable crystalline salt forms of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)-cyclopropanaminefor a method in mono therapy or combing with chemotherapy agents,selected from platinum based or taxane based agents in treating solidtumors, selected from lung, renal, colorectal, gastric, melanoma,head/neck, thyroid, pancreatic, liver, prostate, bladder, brain,sarcoma, breast, ovarian, cervical and endometrial cancers; and bloodcancers, selected from ALL, CLL, AML, CML and Multiple Myeloma;

The present invention relates to bishydrochloride acid,bishydrochloridehydrate acid, bismaleic acid and succinic acid salt, andtheir stable crystalline salt forms or stable crystalline free base formof1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)-cyclopropanaminefor a methods in mono therapy or combining with chemotherapy agents,selected from platinum based or taxane based agents in treating solidtumors, selected from lung, renal, colorectal, gastric, melanoma,head/neck, thyroid, pancreatic, liver, prostate, bladder, brain,sarcoma, breast, ovarian, cervical and endometrial cancers; and bloodcancers, selected from ALL, CLL, AML, CML and Multiple Myeloma;

The present invention relates to bishydrochloride acid,bishydrochloridehydrate acid, bismaleic acid and succinic acid salt, andtheir stable crystalline salt forms or stable crystalline free base formof1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)-cyclopropanaminefor a method in mono therapy or combining with chemotherapy agents,selected from platinum based or taxane based agents in treating lung,colorectal, gastric, thyroid, pancreatic, liver, prostate, sarcoma,breast, ovarian, cervical and endometrial cancers.

The present invention relates to bishydrochloride acid,bishydrochloridehydrate acid, bismaleic acid and succinic acid salt, andtheir stable crystalline salt forms or stable crystalline free base formof1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)-cyclopropanaminefor a method in combining with immunotherapy agents, selected from PD-1or PD-L1, SLAM7, oncolytic virus therapy, bispecific T cell engagers(BiTE) and chimeric antigen receptor (CAR) T cell therapy based agents,such as nivolumab, pembrolizumab, ipilimumab, blinatumomab, elotuzumab,daratumumab, talimogene laherparepvec, in treating solid tumors,selected from lung, renal, colorectal, gastric, melanoma, head/neck,thyroid, pancreatic, liver, prostate, bladder, brain, sarcoma, breast,ovarian, cervical and endometrial cancers; and blood cancers, selectedfrom ALL, CLL, AML, CML and Multiple Myeloma;

The present invention relates a new process to synthesize1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine(AL3818) by condensing intermediate (X1) with (Y1) in a solvent at thepresence of KI or NaI to form intermediate (Z) which is deprotected togive the final compound (AL3818) according to Process A.

The final compound (AL3818) was prepared according to Process A1 when Ris H by deprotecting intermediate (Z-1) with HCOONH₄ (ammonium formate)and Pd/C in an alcoholic solvent, such as MeOH, at 25° C.-80° C. for0.1-4 hours. (Z-1) was prepared by reacting intermediate (X1) with(Y1-1) at the presence of KI or NaI with K₂CO₃ in a solvent, such asacetone or DMF, at a temperature of 60° C.-160° C. for 2-24 hours.

The final compound (AL3818) was prepared according to Process A2 when Ris 4-OMe by deprotecting intermediate (Z-2) with TFA in DCM at 0° C.-30°C. for 1-24 hours. (Z-2) was prepared by reacting intermediate (X1) with(Y1-2) at the presence of KI or NaI with K₂CO₃ in a solvent, such asacetone or DMF, at a temperature of 60° C.-160° C. for 2-24 hours.

The present invention relates a new process to synthesize1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropanamine(AL3818) by reacting intermediate (X2) with (Y2) in a solvent to formintermediate (Z) which is deprotected to give the final compound(AL3818) according to Process B.

The final compound (AL3818) was prepared according to Process B1 when Ris H by deprotecting intermediate (Z-1) with HCOONH₄ (ammonium formate)and Pd/C in an alcoholic solvent, such as MeOH, at 25° C.-80° C. for0.1-4 hours. (Z-1) was prepared by reacting intermediate (X2-1) with(Y2) in a solvent, such as pyridine or lutidine, at a temperature of 60°C.-160° C. for 1-12 hours.

The final compound (AL3818) was prepared according to Process B2 when Ris 4-OMe by deprotecting intermediate (Z-2) with TFA in DCM at 0° C.-30°C. for 1-24 hours. (Z-2) was prepared by reacting intermediate (X2-2)with (Y2) in a solvent, such as pyridine or lutidine, at a temperatureof 60° C.-160° C. for 1-12 hours.

The following examples further illustrate the present invention, butshould not be construed as in any way to limit its scope.

Example 1

Representation of Process A, Process A1

Process for Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)methyl)cyclopropanamine(AL3818)

To a stirred mixture of benzyl 1-(hydroxymethyl)cyclopropylcarbamate (50g) and DCM (200 ml) was added DIPEA (39 g). The result solution wascooled to 0-5° C. with ice/water and further stirred under thistemperature for 15 min. MsCl (30 g) was added via an addition funneldropwise keeping temperature below 5° C. for about 1.5 hours. Aftercompletion of addition, the reaction mixture was allowed stirring at0-5° C. for 30 min and quenched with saturated NaHCO₃ (150 ml). Thesolution was extracted with 150 ml DCM twice. The combined DCM layer waswashed with 0.1 N HCl (400 ml) followed by brine. It was dried overNa₂SO₄ and concentrated to obtain an off-white solid 60 gram as(1-(benzyloxycarbonylamino)cyclopropyl)methyl methanesulfonate (Y1-1),MS: (M+1) 300.

To a stirred mixture of (Y1-1) (16 g), X1[(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-7-hydroxyquinoline,12 g], K₂CO₃ (21 g) and KI (21 g) was added DMF (100 ml), the reactionsuspension was heated at 80° C. for 10 hours and (Y1-1) (10 g) was addedto continuously heated 80° C. for 10 hours. The reaction then wasquenched with water (150 ml) and extracted with 150 ml DCM twice. Thecombined DCM layer was washed with 2 N NaOH (100 ml) followed by waterand brine. It was dried over Na₂SO₄ and concentrated, furtherrecrystallized from EtOH to obtain a yellow solid as benzyl1-((4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)methyl)cyclopropylcarbamate(Z-1) 9.5 g. MS: (M+1) 542.

To a stirred mixture of (Z-1) (9.5 g), HCOONH₄ (4.7 g) and Pd/C (10%,wet 50%, 4.7 g)

was added MeOH, the reaction mixture was heated at 45° C. for 1.5 hours.It was then cooled and filtered through Celite, further evaporated. 2NHCl (200 ml) was added and extracted with DCM/MeOH (10/1, 100 ml) twice.The aqueous layer was basified with 3N NaOH to adjust pH 11-12 togenerate a solid precipitation. The solid was filtered and washed withwater to neutral, further suction dry. The solid was dissolved into amixture of DCM/MeOH (250 ml, 10/1) and further washed with water andbrine. It was dried with MgSO₄ and filtered, further evaporated to givea light yellow solid 5.5 g crude product. Further purification wasconducted by dissolving the crude product into DCM/MeOH (40 ml, 10/1) totriturate with petroleum ether (40 ml) for 2 hours slow stirring. Theprecipitate was filtered and dried in an oven to give the finalcrystalline product 4.4 g (MP: 203-208° C.) and it can be furtherpurified by recrystallizing from EtOH to give purer final product as asame crystalline form. MS: (M+1) 408; ¹H NMR (DMSO-d₆) δ 0.60-0.63 (d,4H), 2.41 (s, 1H), 2.42-2.51 (t, 2H), 3.31 (s, 2H), 3.96 (s, 3H), 4.04(s, 2H), 6.27 (s, 1H), 6.31-6.32 (m, 1H), 6.97-7.02 (t, 1H), 7.20-7.22(d, 1H), 7.36 (s, 1H), 7.60 (s, 1H), 8.40-8.42 (d, 1H), 11.41 (s, 1H).MP: 208-210° C.; DSC Melting Range (Endo): 207-220° C. with PeakTemp=216° C. TGA demonstrating as an unsolvated material with weightloss at about 210° C. (between 205-215° C.). XRPD having patterncompromising characteristic 10 peaks with intensity % greater than 10%expressed in d values and angles as follows:

Angle d value 13.344 6.62986 15.858 5.58405 16.799 5.27326 17.6405.02377 18.770 4.72373 20.650 4.29771 21.633 4.10463 23.087 3.8493425.128 3.54112 26.607 3.34755XRPD having pattern compromising 26 characteristic peaks with allintensity % expressed in d values and angles as follows:

NO. Angle d value Intensity (%) 1 10.892 8.11623 2.1 2 11.589 7.629916.1 3 12.195 7.25174 5.9 4 13.344 6.62986 36.2 5 15.858 5.58405 31.5 616.799 5.27326 77.9 7 17.640 5.02377 18.8 8 18.770 4.72373 11.9 9 19.9874.43884 7.2 10 20.650 4.29771 42.0 11 21.633 4.10463 15.3 12 23.0873.84934 100.0 13 24.356 3.65157 3.5 14 25.128 3.54112 14.6 15 25.6693.46768 3.8 16 26.607 3.34755 18.0 17 26.607 3.34755 3.1 18 29.0503.07132 5.7 19 29.797 2.99602 1.5 20 30.681 2.91167 4.3 21 31.8532.80718 1.2 22 33.524 2.67095 2.8 23 34.789 2.57667 2.6 24 35.8732.50131 2.2 25 37.391 2.40313 3.9 26 38.637 2.32846 1.4Graphs of DSC, TGA and XRPD are represented by FIG. 1, FIG. 2 and FIG. 3respectively.

Example 2

Representation of Process A, Process A2

Process for Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)methyl)cyclopropanamine(AL3818)

It was similar prepared according to the preparation procedures of (Z-1)described in Example 1 by using 4-methoxybenzyl1-(hydroxymethyl)cyclopropylcarbamate to first generate(1-((4-methoxybenzyloxy)carbonylamino)cyclopropyl)methylmethanesulfonate (Y1-2) then to give 4-methoxybenzyl1-((4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)-methyl)cyclopropylcarbamate(Z-2), MS: (M+1) 572

To a stirred mixture of (Z-2) (1.5 g) in DCM (15 ml) at 0° C. was addedTFA (1.5 ml) for about 30 min and warmed up to RT. The reaction wasstirred at RT for 2 hours and added into water (30 ml). The aqueouslayer was extracted with DCM twice (100 ml×2) and basified with 2N NaOHto adjust pH 11-12. The mixture was extracted with DCM (100 ml×3) andfurther washed with brine (100 ml). It was dried with MgSO₄ andfiltered. The solution was evaporated to give 1.05 g crude finalproduct. Further purification was conducted to dissolve the crudeproduct into DCM/MeOH and triturated with petroleum ether and dried inan oven to give the final pure product 0.8 g AL3818 with the samecrystalline form.

Example 3

Representation of Process A, Process B1

Process for Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)methyl)cyclopropanamine(AL3818)

To a mixture of benzyl1-((4-chloro-6-methoxyquinolin-7-yloxy)methyl)cyclopropyl-carbamate(X2-1) (5 g), 4-fluoro-2-methyl-1H-indol-5-ol (Y2) (5 g) and DMAP (4 g)was added 1,6-lutidine (15 ml). The reaction was stirred and heated at135° C. for 5 hours and was cooled followed by adding IPA with slowstirring for 2 hours at RT. The solid was filtered and further washedwith IPA, dried to give (Z-1) 5.2 g as a solid. It was then similarlyprepared according to deprotection procedures described of (Z-1) inExample 1 to give the final compound AL3818 with the same crystallineform.

Example 4

Representation of Process A, Process B2

Process for Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)methyl)cyclopropanamine(AL3818)

(Z-2) was similarly prepared according to the procedures described inExample 3 by using 4-methoxybenzyl1-((4-chloro-6-methoxyquinolin-7-yloxy)methyl)cyclopropylcarbamate(X2-2) and (Y2). It was then similarly prepared according todeprotection procedures of (Z-2) described in Example 2 to give thefinal compound AL3818 with the same crystalline form.

Example 5 Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)-methyl)cyclopropanamineBishydrochloride Acid Salt and its Crystalline

To a 25 ml flask was added 250 mg free base (AL3818), 4N HCl in dioxane0.625 mL (2.5 mmol, 4 eq.) in 10 ml EtOH, the reaction was heated at 75°C. for 30 minutes, cooled to RT and stirred for O.N. The solid wasfiltered and rinsed with acetone twice. It was dried in oven at 50° C.for 4 hours to give 126 mg white solid as the bishydrochloride salt as acrystalline and further recrystallized from EtOH to give a purer productas a same crystalline form. ¹H NMR (DMSO-d₆) δ 1.09-1.24 (m, 4H), 2.43(s, 3H), 4.08 (s, 3H), 4.40 (s, 2H), 6.32 (s, 1H), 6.76 (s, 1H),7.05-7.11 (t, 1H), 7.27-7.30 (d, 1H), 7.65 (s, 1H), 7.82 (s, 1H), 8.64(s, 2H), 8.70-8.73 (m, 1H), 11.51 (s, 1H). Chloride ion chromatographyshowed 2 molecular ratio ions (16.1%). DSC Melting Range (Exo): 249-280with Peak Temp=268° C. TGA demonstrating as an unsolvated material withweight loss at about 230° C. (between 225-235° C.). XRPD having patterncompromising 21 characteristic peaks with intensity % greater than 10%or 27 peaks with all intensity % expressed in d values and angles asfollows:

NO. Angle d value Intensity (%) 1 7.640 11.56173 19.5 2 8.642 10.2232820 3 9.361 9.43969 13.3 4 10.091 8.75881 100.0 5 13.740 6.43957 26.4 614.479 6.11252 54.7 7 15.186 5.82962 10.1 8 15.766 5.61643 20.3 9 17.2065.14957 7.4 10 18.569 4.77448 18.6 11 19.271 4.60215 11.0 12 20.0414.42696 49.5 13 22.211 3.99909 58.4 14 22.814 3.89483 11.2 15 23.3983.79886 11.6 16 24.455 3.63702 76.6 17 25.524 3.48708 34.6 18 26.7033.33576 21.7 19 27.337 3.25978 18.4 20 28.061 3.17732 18.5 21 28.8013.09732 6.3 22 29.845 2.99133 13.8 23 31.331 2.85271 7.1 24 31.6212.82721 9.5 25 32.840 2.72504 10.5 26 33.714 2.65632 3.8 27 38.3482.34534 9.6Graphs of DSC, TGA and XRPD are represented by FIG. 4, FIG. 5 and FIG. 6respectively.

Example 6 Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)methyl)cyclopropanaminebishydrochloridehydrate Acid Salt and its Crystalline

To a 10 mL flask, charged 140 mg of 3818-2HCl salt from above Example 4and 0.7 mL (×5 with salt volume) of 80% MeOH in H₂O. The resultsuspension was heated to 70° C. to form a solution and cooled to RT andfurther stirred for O.N. The solid was filtered and rinsed with acetonetwice. It was dried in oven at 50° C. for 4 hours to obtain off-whitesolid 110 mg as the crystalline bishydrochloride hydrate salt. ¹H NMR(DMSO-d₆) δ 1.09 (s, 2H), 1.22 (s, 2H), 2.44 (s, 1H), 2.52 (s, 2H), 4.09(s, 3H), 4.44 (s, 2H), 6.32 (s, 1H), 6.81-6.82 (d, 1H), 7.08-7.14 (t,1H), 7.29-7.32 (d, 1H), 7.79 (s, 1H), 7.85 (s, 1H), 8.75-8.78 (d, 1H),8.85 (s, 2H), 11.66 (s. 1H). Chloride ion chromatography showed 2molecular ratio ions (17.8%). DSC Melting Range (Exo): 207-260° C. withPeak Temp=226° C. TGA demonstrating 2.68% (˜3%, 1 water) weight losstill 120° C. (between 115-125° C.) and further weight loss at about 170°C. (between 165-175° C.). XRPD having pattern comprising 9characteristic peaks with intensity % greater than 10% or 12 peaks withall intensity % expressed in d values and angles as follows:

NO. Angle d value Intensity (%) 1 5.506 16.03679 28.0 2 6.817 12.95694100 3 8.087 10.92445 29.9 4 9.766 9.04936 20.6 5 13.318 6.64283 22.3 614.332 6.17523 7.0 7 16.159 5.48067 15.7 8 19.474 4.55451 8.8 9 20.9204.24296 6.5 10 20.920 3.87231 28.2 11 25.087 3.54678 20.2 12 25.8743.44064 22.7Graphs of DSC, TGA and XRPD are represented by FIG. 7, FIG. 8 and FIG. 9respectively.

Example 7 Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)-methyl)cyclopropanamineBismaleic Acid Salt and its Crystalline

To a 25 mL flask was added 50 mg free base (AL3818) in 1.5 mL EtOH, thereaction was heated to 70° C. with stirring. To the result solution wasadded 36 mg (2.5 eq) maleic acid as solid and stirred at 70° C. for 0.5hr. It was cooled to RT and stirred for O.N. The solid was filtered andrinsed with acetone twice further dried in oven at 50° C. for 4 hours toobtain 68 mg of as a crystalline solid and further recrystallized fromEtOH to give a purer product as a same crystalline form as two (bis)maleic acid salt. ¹H NMR (DMSO-d₆) δ 0.73 (s, 2H), 0.88 (s, 2H), 3.43(s, 2H), 3.53 (s, 2H), 3.59 (s, 2H), 3.86 (s, 4H), 3.97 (s, 3H), 4.41(s, 1H), 6.07 (s, 2H), 7.26 (s, 1H), 7.44-7.50 (t, 1H), 7.76-7.79 (d,1H), 7.88 (s, 1H), 8.10-8.12 (d, 1H), 8.55 (s, 1H), 9.54 (s. 1H). Maleicion chromatography showed 2 molecular ratio ions (37.1%). DSC MeltingRange (Endo): 165-202° C. with Peak Temp=183° C. TGA demonstrating as anunsolvated material with weight loss at about 160° C. (between 155-165°C.). XRPD having pattern comprising 22 characteristic peaks withintensity % greater than 10% or 35 peaks with all intensity % expressedin d values and angles as follows:

NO. Angle d value Intensity (%) 1 6.716 13.14986 29.7 2 8.816 10.0218934.3 3 9.743 9.07069 15.3 4 10.033 8.80923 21.4 5 11.777 7.50803 21.2 613.418 6.59342 6.2 7 14.816 5.97445 11.0 8 16.089 5.50434 9.5 9 16.8015.27279 24.5 10 17.360 5.10409 87.9 11 17.179 5.15755 70.7 12 18.1904.87308 20.2 13 18.704 4.74028 16.7 14 19.296 4.59623 5.0 15 19.9204.45371 12.6 16 20.824 4.26227 65.5 17 21.457 4.13785 100.0 18 22.4113.96393 4.5 19 22.876 3.88434 5.8 20 23.204 3.83021 19.0 21 23.6223.76332 78.4 22 24.418 3.64247 6.3 23 26.140 3.40621 87.0 24 26.9583.30469 26.5 25 27.383 3.25443 61.3 26 28.154 3.16697 41.5 27 29.5543.02013 6.8 28 30.611 2.91815 23.7 29 31.373 2.84906 14.3 30 33.4572.67620 6.7 31 34.541 2.59465 2.8 32 35.137 2.55199 3.8 33 35.7342.51067 2.5 34 37.129 2.41949 8.6 35 39.704 2.26833 3.9Graphs of DSC, TGA and XRPD are represented by FIG. 10, FIG. 11 and FIG.12 respectively.

Example 8 Preparation of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quinolin-7-yloxy)methyl)cyclopropanamineSuccinic Acid Salt and its Crystalline

To a 50 mL flask was added 100 mg free base (AL3818) in 4 mL EtOH, thereaction was heated to 75° C. with stirring. To the result solution wasadded succinic acid as solid 36 mg (0.308 mmol, 1.25 eq) and stirred at75° C. for 0.5 hr. It was cooled to RT and stirred for overnight. Thesolid was filtered and rinsed with acetone twice further dried in ovenat 50° C. for 4 hours to obtain a crystalline solid 84 mg and furtherrecrystallized from EtOH to give a purer product as a same crystallineform. ¹H NMR (DMSO-d₆) δ 0.72 (s, 4H), 2.37-2.42 (m, 7H), 3.99 (s, 3H),4.10 (s, 2H), 6.27 (s, 1H), 6.32-6.33 (d, 1H), 6.97-7.02 (t, 1H),7.20-7.23 (d, 1H), 7.39 (s, 1H), 7.61 (s, 1H), 8.42 (s, 2H), 11.41 (s,1H). Succinic ion chromatography showed 1 molecular ratio ions (23.2%).DSC Melting Range: Melting Range (Endo): 176-202° C. with Peak Temp=198°C. TGA demonstrating as an unsolvated material with weight loss at about180° C. (between 175-185° C.). XRPD having pattern comprising 11characteristic peaks with intensity % greater than 10% or 19 peaks withall intensity %:

NO. Angle d value Intensity (%) 1  5.765 15.31849 12.9 2  8.038 10.989947.4 3 11.639 7.59700 27.7 4  12.950 6 6.83065 100 5  16.141 5 5.4868318.0 6  17.483 5 5.06846 18.7 7 18.385 4.82175 17.8 8 19.394 4.57325 1.19 20.756 4.27609 13.4 10 22.034 4.03092 2.8 11 23.167 3.83630 1.8 1224.085 3.69200 16.9 13 24.485 3.63268 14.6 14 25.737 3.45874 13.7 1528.621 3.11637 6.4 16 29.255 3.05025 22.1 17 31.357 2.85048 0.9 1831.967 2.79743 2.1 19 35.630 2.51780 2.4Graphs of DSC, TGA and XRPD are represented by FIG. 13, FIG. 14 and FIG.15 respectively.

Example 9

In vitro MTT (proliferation) assay was performed with compound fromabove examples to give following inhibition results:

Sample Name AL3818-H1 AL3818-H2 AL3818-S AL3818-M AL3818-F (2HCl)(2HCl•HCl) (Succinic) (Bismaleic) (Free Base) Cancer Cell Lines IC50(μM) IC50 (μM) IC50 (μM) IC50 (μM) IC50 (μM) PANC-1 (pancreatic) 1.581.55 1.24 1.15 1.34 NC1-H157 (lung) 7.18 2.29 2.28 2.02 1.83 MDA-MB-23110.25 5.71 6.1 5.37 5.56 (breast) Hela (cervical) 3.7 4.07 4.66 3.943.23 PC-3 (prostate) 5.53 3.88 4.41 3.62 5.11 BEL7404 (liver) 2.75 1.741.43 1.15 0.93 MKN45 (gastric) 1.77 3.9 4.16 2.65 3.11 Ishikawa 5.671.56 1.32 1.34 1.04 (endometrial) Saos-2 (sarcoma) 4.68 4.98 5.68 5.495.13 SKOV3 (ovarian) 5.34 5.7 7.6 6.56 5.97 SW579 (thyroid) 2.33 1.292.61 1.15 1 HCT1 16 (colon) 3.45 3.34 3.94 4.62 3.25

Example 10

Based on the inventor's research experience using AL3818 free base, its2HCl salts, its bis-maleic acid salt and its succinic salt, thefollowing tumor inhibition results are expected in a MTT assay accordingto Example 9.

2-10 μM±1.7 μM in vitro inhibition activities are expected on varioussolid tumor cell lines, such as renal, melanoma, head/neck, bladder,brain; and blood cancers, such as ALL, CLL, AML, CML and MultipleMyeloma.

Example 11

Animal antitumor activity in vivo testing with endometrial Ishikawa cellline (xenograft) is performed as follows:

The well grown tumor tissue of endometrial cancer Ishikawa was cut into3 mm pieces, and each nude mouse was subcutaneously inoculated with onepiece into the right armpit. The animals were grouped and administratedas following:

-   -   1) AL3818-H1 (Bishydrochloride acid salt, 2HCl), MW: 480, 3.54        mg/kg    -   2) AL3818-H2 (Bishydrochloridehydrate acid salt, 2HCl.H₂O), MW:        598, 3.67 mg/kg    -   3) AL3818-S(Succinic acid salt), MW: 525, 3.87 mg/kg    -   4) AL3818-M (Bismaleic acid salt), MW: 639, 4.71 mg/kg    -   5) AL3818-F (Free Base), MW: 407, 3 mg/kg    -   6) Control        Treatments were initiated when the tumors size reached above 100        mm³ after 13 days. According to the size of tumor, the animals        with oversize or undersize tumors were eliminated, and the        animals were grouped with similar average tumor volume. Then the        animals were oral administrated daily for continuous 14 days        with volume of 0.5 ml/20 g as the above. The large diameter        a (mm) and the small diameter b (mm) were measured with caliper        twice a week after inoculation for 13 days. The tumor volume was        calculated by formular: TV=ab²/2. The relative tumor volume was        calculated as: RTV=Vt/Vo, Vo represents the tumor volume on the        first day of treatment; Vt represents the tumor volume on each        measurement day. The animals were executed and tumors were got        by dissection 30 days after inoculation (D18). Then, the        individual body weight and tumor weight were determined, and        calculate as following formula.        Tumor inhibition activities are between 50-95%. Results are        shown in FIG. 16.

Example 12

Based on the inventor's research experience using AL3818 free base, itsHCl salts (mono or bis), its bis-maleic acid salt and its succinic salt,the following in vivo tumor inhibition results (xenografts) are expectedaccording to Example 11.

50%-100% in vivo tumor inhibition activities are expected on varioussolid tumor cell lines, such as lung, renal, colorectal, gastric,melanoma, head/neck, thyroid, pancreatic, liver, prostate, bladder,brain, sarcoma, breast, ovarian, cervical and endometrial cancers; andblood cancers, such as ALL, CLL, AML, CML and Multiple Myeloma.

Example 13

AL3818 bis HCl salt and bis-maleic acid salt were also tested incombining with chemotherapy by using platinum based, taxane based orboth; such as: cisplatin, carboplatin, paclitaxel orcisplatin/paclitaxel, carboplatin/paclitaxel combined. The experiment ofcombination with carboplatin/paclitaxel was carried out similar to thedescription of Example 11. Tumor inhibition activities are between 50to >100%. Results are shown in FIG. 17.

Example 14

Based on the inventor's research experience using AL3818 free base, itsHCl salts (mono or bis), its bis-maleic acid salt and its succinic salt,the following in vivo combining chemotherapy (standard of care, such asplatinum based, taxane based or both chemotherapy) tumor inhibitionresults (xenografts) are expected according to Example 13, especiallycombining with cisplatin, carboplatin, paclitaxel orcisplatin/paclitaxel, carboplatin/paclitaxel together.50 to >100% regression in vivo tumor inhibition activities are expectedon various solid tumor cell lines, such as lung, renal, colorectal,gastric, melanoma, head/neck, thyroid, pancreatic, liver, prostate,bladder, brain, sarcoma, breast, ovarian and cervical cancers; and bloodcancers, such as ALL, CLL, AML, CML and Multiple Myeloma.

Example 15

Based on the inventor's research experience using AL3818 free base, itsHCl salts (mono or bis), its bis-maleic acid salt and its succinic salt,the following in vivo combining effects are expected, which in combiningwith immunotherapy agents, selected from PD-1 or PD-L1, SLAM7, oncolyticvirus therapy, bispecific T cell engagers (BiTE) and chimeric antigenreceptor (CAR) T cell therapy based agents, such as nivolumab,pembrolizumab, ipilimumab, blinatumomab, elotuzumab, daratumumab,talimogene laherparepvec but not limited, in treating solid tumors,selected from lung, renal, colorectal, gastric, melanoma, head/neck,thyroid, pancreatic, liver, prostate, bladder, brain, sarcoma, breast,ovarian, cervical and endometrial cancers; and blood cancers, selectedfrom ALL, CLL, AML, CML and Multiple Myeloma50 to >100% regression in vivo tumor inhibition activities are expectedon various solid tumor cell lines, such as lung, renal, colorectal,gastric, melanoma, head/neck, thyroid, pancreatic, liver, prostate,bladder, brain, sarcoma, breast, ovarian and cervical cancers; and bloodcancers, such as ALL, CLL, AML, CML and Multiple Myeloma.

Example 16

A mouse model of laser-induced choroidal neovascularization (CNV)

(1) Experiments were performed in C57 BL/6 mice at 10 to 12 weeks ofage. Laser CNV was induced with a 532 nm diode laser mounted on a slitlamp, using 50 m spot size, 100 ms duration and 100 mW laser energy.Each eye received 4 laser burns. Stock solution of AL3818-H1 was made bydissolving the compound in water to 25 mg/ml concentration, and furtherdiluted to working solution of either 250 or 25 μg/ml in water. Micewere orally gavaged at dosage of 2.5 or 0.25 mg/kg body weight, involume of 200 μl per 20 gram of body weight, once every day starting atone day before laser treatment until 10 days afterwards. Control groupmice will receive gavage with water which is used to dissolve thecompound. By the end of the experiment, mice were subjected tofluorescence angiography to exclude the spots with hemorrhage and othermechanic injuries caused by the procedures. Mice were sacrificed and CNVsize was measured by immunostaining of RPE/choroid flat mount co-stainedwith FITC-conjugated isolectin B4 and anti-ICAM2 antibody. For groups ofcontrol, 0.25 mg/kg and 2.5 mg/kg, we examined 43, 44 and 49 eyes onRPE/choroid flat mount. After immunostaining, images were taken on aZeiss fluorescence microscope. CNV size was measured in Image Jsoftware. Our results (FIG. 18) show that mice treated with AL3818 at0.25 or 2.5 mg/kg body weight had nearly 70% reduction in average sizeof laser-induced CNV.(2) The potential synergistic or additive effects between AL3818-M andanti-VEGF antibody was also studied. Immediately after laser burn, micewere treated with a monoclonal VEGF neutralizing antibody from R&DSystems (mAb AF564) at 1 μg dosage by intravitreal injection. We had 4experimental groups: control (treated with water), AL3818-M, anti-VEGF,and AL3818-M+anti-VEGF. A total of 75 laser spots were analyzed. Mice incontrol or AL3818-M alone group received the intravitreal injection ofsaline in the same 2 μl volume. The results ((FIG. 19) showed that eyestreated with 0.15 mg/kg AL3818 and 1 μg anti-VEGF antibody had a nearly30% reduction in laser CNV as compared to the control group (P<0.01,one-way ANOVA, Dunnett post-hoc test).

Example 17

Based on the inventor's research experience using AL3818 free base, itsHCl salts (mono or bis), its bis-maleic acid salt and its succinic salt,in vivo animal model CNV efficacies which related to effective treatmentof an optometric disease, such as AMD (Age-Related Macular Degeneration)but not limited, are definitely expected according to Example 16; orcombing these compounds with anti-VEGF antibody or VEGF trap, such asranibizumab or aflibercep but not limited, to generate a synergistictreatment effect are definitely expected according to Example 16 aswell.

Example 18

Conventional water solubility test has been conducted to give thefollowing results:

Sample Name AL3818-H1 AL3818-H2 AL3818-S AL3818-M AL3818-F (2HCl)(2HCl•H2O) (Succinic) (2Maleic) (Free Base) (mg/ml) (mg/ml) (mg/ml)(mg/ml) (mg/ml) water solubility 6 7 0.1 0.5 0.02

What is claimed is:
 1. A crystalline form of a compound,1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quino-lin-7-yloxy)methyl)cyclopropanamine,wherein the crystalline form exhibits a XRPD having the followingcharacteristic peaks: NO. Angle 1 7.640 2 8.642 3 9.361 4 10.091 513.740 6 14.479 7 15.186 8 15.766 9 17.206 10 18.569 11 19.271 12 20.04113 22.211 14 22.814 15 23.398 16 24.455 17 25.524 18 26.703 19 27.337 2028.061 21 28.801 22 29.845 23 31.331 24 31.621 25 32.840 26 33.714 2738.348.


2. The compound of claim 1, wherein the characteristic peaks have anintensity greater than about 10%.
 3. The compound of claim 1, whereinthe peaks have d values of: NO. d value 1 11.56173 2 10.22328 3 9.439694 8.75881 5 6.43957 6 6.11252 7 5.82962 8 5.61643 9 5.14957 10 4.7744811 4.60215 12 4.42696 13 3.99909 14 3.89483 15 3.79886 16 3.63702 173.48708 18 3.33576 19 3.25978 20 3.17732 21 3.09732 22 2.99133 232.85271 24 2.82721 25 2.72504 26 2.65632 27 2.34534.


4. The compound of claim 1, wherein the peaks have intensity values asfollows: NO. Intensity (%) 1 19.5 2 20 3 13.3 4 100.0 5 26.4 6 54.7 710.1 8 20.3 9 7.4 10 18.6 11 11.0 12 49.5 13 58.4 14 11.2 15 11.6 1676.6 17 34.6 18 12.7 19 18.4 20 18.5 21 6.3 22 13.8 23 7.1 24 9.5 2510.5 26 3.8 27 9.6.


5. The compound of claim 1, wherein the1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quino-lin-7-yloxy)methyl)cyclopropanamineis a bishydrochloride salt.
 6. A method of treating a neoplasticdisease, said method comprising: administering to a patient in needthereof a salt of1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquino-lin-7-yloxy)-methyl)-cyclopropanamine;and administering a second therapeutic agent to the patient; wherein the1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquino-lin-7-yloxy)-methyl)-cyclopropanaminesalt is selected from: a bishydrochloride acid salt, abishydrochloridehydrate acid salt, a bismaleic acid salt and a succinicacid salt; wherein the neoplastic disease comprises solid tumors,selected from lung, renal, colorectal, gastric, melanoma, head/neck,thyroid, pancreatic, liver, prostate, bladder, brain, sarcoma, breast,ovarian, cervical and endometrial cancers; and blood cancers, selectedfrom ALL, CLL, AML, CML and Multiple Myeloma.
 7. The method of claim 6,wherein the second therapeutic agent is a chemotherapeutic agent.
 8. Themethod of claim 7, wherein the second therapeutic agent is selected froma platinum-based agent and a taxane-based agents.
 9. The method of claim7, wherein the second therapeutic agent is selected from paclitaxel,cisplatin, and carboplatin.
 10. The method of claim 7, furthercomprising administering a third therapeutic agent to the patient. 11.The method of claim 10, wherein the second therapeutic agent ispaclitaxel and the third therapeutic agent is cisplatin or carboplatin.12. The method of claim 9, further comprising administering a thirdtherapeutic agent to the patient wherein the third therapeutic agent isan immunotherapy agent.
 13. The method of claim 12, wherein theimmunotherapy agent is selected from nivolumab, pembrolizumab,ipilimumab, blinatumomab, elotuzumab, daratumumab, and a talimogenelaherparepvec based agent.
 14. The method of claim 6, wherein the secondtherapeutic agent is an immunotherapy agent.
 15. The method of claim 14,wherein the immunotherapy agent is selected from nivolumab,pembrolizumab, ipilimumab, blinatumomab, elotuzumab, daratumumab, and atalimogene laherparepvec based agent.
 16. The method of claim 6, whereinthe second therapeutic agent is an anti-VEGF antibody or a VEGF trap.17. A crystalline form of a compound,1-((4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxy-quino-lin-7-yloxy)methyl)cyclopropanamine,wherein the crystalline form exhibits a XRPD having the followingcharacteristic peaks: NO. Angle 1 10.091 2 14.479 3 20.041 4 22.211 524.455.


18. The compound of claim 17, wherein the characteristic peaks have anintensity greater than about 10%.
 19. A method of treating a neoplasticdisease, said method comprising: administering the compound of claim 17and a second therapeutic agent to a patient in need thereof; and whereinthe neoplastic disease comprises solid tumors, selected from lung,renal, colorectal, gastric, melanoma, head/neck, thyroid, pancreatic,liver, prostate, bladder, brain, sarcoma, breast, ovarian, cervical andendometrial cancers; and blood cancers, selected from ALL, CLL, AML, CMLand Multiple Myeloma.
 20. The method of claim 19, further comprisingadministering a third therapeutic agent to the patient.